DE10043667A1 - 2-guanidino-4-aryl-quinazolines - Google Patents

Abstract

The invention relates to compounds of formula (I), wherein Y represents (II) or (III) and Ar, R<1>, R<2>, R<5>, R<6>, R<7> and R<8> have the above-mentioned meaning, in addition to the salts and solvates thereof and the use thereof as NHE-3-inhibitors.

Description

The invention relates to compounds of the formula I.

Ar unsubstituted or simply substituted by R ³ and / or R ⁴ phenyl or naphthyl,
R ¹ , R ² , R ³ , R ⁴ each independently of one another H, A, OA, Hal, CF ₃ , OH, NO ₂ , NH ₂ , NHA, NA ₂ , NH-CO-A, NH-CO-Ph, SA, SO-A, SO ₂ -A, SO ₂ - Ph, CN, OCF ₃ , CO-A, CO ₂ H, CO ₂ A, CO-NH ₂ , CO-NHA, CO-NA ₂ , SO ₂ NH ₂ , SO ₂ NHA, SO ₂ NA ₂ or unsubstituted or mono- or polysubstituted by A, OA, Hal or CF ₃ ,
A alkyl having 1, 2, 3, 4, 5 or 6 carbon atoms,
Hal F, Cl, Br or I,
R ⁵ , R ⁶ , R ⁷ , R ⁸ each independently represent H, A, unsubstituted or mono- or polysubstituted by A, OA, Hal, CF ₃ , where R ⁵ and R ⁷ , R ⁵ and R ⁶ , R ⁷ and R ⁸ can form 5-7-membered rings,
and their salts and solvates, with the proviso that compounds in which R ⁵ , R ⁶ , R ⁷ and R ^{8 are} simultaneously H and none of the radicals R ¹ , R ² , R ³ , R ⁴ OH, NO ₂ , NH ₂ , NHA, NA ₂ , NH-CO-A, NH-CO-Ph, SA, SO-A, SO ₂ -A, SO ₂ -Ph, CN, OCF ₃ , CO-A, CO ₂ H, CO ₂ A, CO-NH ₂ , CO-NHA, CO-NA ₂ , SO ₂ NH ₂ , SO ₂ NHA, SO ₂ NA ₂ or unsubstituted or mono- or polysubstituted by A, OA, Hal or CF ₃ , except are.

The invention also relates to the use of the compounds of Formula I and its salts and solvates as NHE-3 inhibitors.

Other inhibitors of the sodium / proton exchanger subtype 3 are e.g. B. in EP 0 825 178.

The exempted connections are already in US 3,131,187 and their use for other purposes. Quinazolinyl guanidine derivatives are described by V.I. Shvedov et al. in Pharm. Chem. J. (Engl. Transl.) 1980, 14, 532-538 or in Khim. Farm. Zh. 1980, 14, 38-43, and by S.C. Bell et al. in J. Med. Pharm. Chem. 1962, 5, 63-69.

The invention was based on the object, new compounds with valuable len properties to find, especially those that are used to manufacture of drugs can be used.

Surprisingly, it has been found that the compounds of the formula I and its salts with good tolerance the sodium / proton off inhibit exchanger subtype 3.

The compounds of formula I can be used as active pharmaceutical ingredients in the Human and veterinary medicine are used.

It is known that the Na ⁺ / H ⁺ exchanger is a family with at least 6 different isoforms (NHE-1 to NHE-6), all of which are already cloned. While the NHE-1 subtype is ubiquitously distributed throughout the body in all tissues, the remaining NHE subtypes are selectively expressed in specific organs such as in the kidney or in the lumen wall and contraluminal wall of the small intestine. This distribution reflects the specific functions to which the various isoforms serve, namely on the one hand the regulation of the intracellular pH and the cell volume by the subtype NHE-1 and on the other hand the Na ⁺ uptake and reuptake in the intestine and kidney by the Isoforms NHE-2 or NHE-3. The NHE-4 isoform was found mainly in the stomach. The expression of NHE-5 is limited to brain and neuron tissue. NHE-6 represents the isoform that forms the sodium proton exchanger in the mitochondria.

The NHE-3 isoform is found particularly in the apical membrane of the proxima len kidney tubules expressed; an NHE-3 inhibitor therefore exercises u. a. a never protective effect.

The therapeutic use of a selective inhibitor for NHE-3 Isoforming is versatile. NHE-3 inhibitors inhibit or reduce Ge weave damage and cell necrosis after pathophysiological hypoxic and ischemic events leading to activation of NHE activity cause like this during renal ischemia or during removal, the transport and reperfusion of a kidney during kidney transplantation is the case.

The compounds of formula I have a cytoprotective effect by over shooting absorption of sodium and water into the cells of with Prevent oxygen-deficient organs.

The compounds of formula I have a hypotensive effect and are suitable as active pharmaceutical ingredients for the treatment of hypertension. Still own they are called diuretics.

The compounds of formula I act alone or in combination with NHE Inhibitors of other subtype specificity are anti-ischemic and can be used are used to protect against thrombosis, atherosclerosis, vascular spasms Organs, e.g. B. kidney and liver, before and during operations, as well as chronic or acute kidney failure.  

They can also be used to treat stroke, of brain edema, ischemia of the nervous system, various forms of shock, e.g. B. allergic, cardiological, hypovolaean or bacterial shocks, as well as to improve respiratory drive for example the following conditions: central sleep apneas, sudden Child death, postoperative hypoxia and other breathing disorders.

By combining it with a carbonic anhydrase inhibitor, the At management activities can be further improved.

The compounds of formula I have an inhibitory effect on proliferation of cells, for example fibroblast cell proliferation and the Pro liferation of the smooth vascular muscle cells and can therefore be used for treatment tion of diseases are used in which the cell proliferation egg is a primary or secondary cause.

The compounds of formula I can be used against diabetes late complications, cancer, fibrotic diseases, endothelial dysfunction, organ hypertrophy and hyperplasia, esp especially for prostate hyperplasia or prostate hypertrophy.

They are also suitable as diagnostics for determining and differentiating of certain forms of hypertension, atherosclerosis, diabetes tes and proliferative diseases.

Since the compounds of formula I also the level of serum lipoprotei ne beneficial influence, they can be used to treat an elevated Blood lipid levels alone or in combination with other medicines be set.

The invention relates to the use of compounds of Formula I according to claim 1 and its physiologically acceptable salts and / or Solvate for the manufacture of a medicament for the treatment of Thrombosis, ischemic conditions of the heart, peripheral and central nervous system and stroke, ischemic conditions peripheral organs and limbs and for the treatment of shockzu stands.

The invention further relates to the use of compounds of the formula I according to claim 1 and its physiologically acceptable sal ze and / or solvates for the manufacture of a medicament for use in  surgical operations and organ transplants and for preservation Storage and storage of grafts for surgical measures.

The invention also relates to the use of compounds of Formula I according to claim 1 and its physiologically acceptable salts and / or Solvate for the manufacture of a medicament for the treatment of Diseases in which cell proliferation is primary or secondary Cause represents, for the treatment or prophylaxis of disorders of the Fat metabolism or disturbed respiratory drive.

The invention also relates to the use of compounds of the formula I according to claim 1 and its physiologically acceptable sal ze and / or solvate for the manufacture of a medicament for treatment from ischemic kidney, ischemic bowel disease or to pro prevention of acute or chronic kidney disease.

Methods for identifying substances that cause sodium / proton Inhibitors of subtype 3 inhibit z. B. described in US 5,871,919.

The compounds of formula I are also for the treatment of bacteria len and parasitic diseases.

For all residues in the compounds of formula I which occur several times, such as B. A applies that their meanings are independent of each other.

Under hydrates and solvates one understands z. B. the hemi-, mono- or Dihydrates, among solvates e.g. B. alcohol addition compounds such. B. with Methanol or ethanol.

In the above formulas, A means alkyl, is linear or branched, and has 1, 2, 3, 4, 5 or 6 carbon atoms. A is preferably methyl, furthermore ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl or tert.- Butyl, also pentyl, 1-, 2- or 3-methylbutyl, 1,1-, 1,2- or 2,2- Dimethylpropyl, 1-ethylpropyl, hexyl, 1-, 2-, 3- or 4-methylpentyl, 1,1-, 1,2-, 1,3-, 2,2-, 2,3- or 3,3-dimethylbutyl, 1- or 2-ethylbutyl, 1-ethyl- 1-methylpropyl, 1-ethyl-2-methylpropyl, 1,1,2- or 1,2,2-trimethylpropyl.  

OA is preferably methoxy, ethoxy, propoxy, or isopropoxy Butoxy.

Hal is preferably F, Cl or Br, but also I, in particular F, Cl or Br.

Above and below, Ph means an unsubstituted phenyl radical, see above unless otherwise stated.

Ar is preferably unsubstituted phenyl or naphthyl, further preferably z. B. by A, fluorine, chlorine, bromine, iodine, methoxy, ethoxy, propoxy, butoxy or CF ₃ monosubstituted phenyl or naphthyl. Ar is particularly preferably unsubstituted or phenyl monosubstituted by A, fluorine, chlorine, bromine, iodine, methoxy, ethoxy, propoxy, butoxy or CF ₃ .

R ⁵ , R ⁶ , R ⁷ and R ^{8 are} preferably simultaneously H or, independently of one another, H or A, which is defined as indicated above.

If R ⁵ and R ⁷ together form a ring, Y preferably adopts one of the following structures:

wherein R ⁶ and R ^{8 have} the meaning given above and n is 1, 2 or 3, preferably 1 or 2.

If R ⁷ and R ⁸ together form a ring, Y preferably adopts one of the following structures:

wherein R ⁵ and R ^{6 have} the meaning given above and n is 1, 2 or 3, preferably 1 or 2.

If R ⁵ and R ⁶ together form a ring, Y preferably adopts one of the following structures:

wherein R ⁷ and R ^{8 have} the meaning given above and n is 1, 2 or 3, preferably 1 or 2.

The invention relates in particular to the compounds of formula I in which at least one of the said radicals has one of the preferred meanings given above and the use thereof. Some preferred groups of compounds can be expressed by the following partial formulas 1a to 1e, which correspond to the formula I and in which the radicals not specified have the meaning given for the formula I, but in which
in Ia R ¹ H, OH, OA, SA or Hal, in particular H, OH, OCH ₃ or CH ₃
means;
in Ib R ¹ H, OH, OA, SA or Hal, in particular H, OH, OCH ₃ or CH ₃
R ² H, Hal, OH, A, NH ₂ , NO ₂ or CN, in particular H, Cl, OH, CH ₃ or NH ₂
mean;
in Ic R ¹ H, OH, OA, SA or Hal, in particular H, OH, OCH ₃ or CH ₃
R ² H, Hal, OH, A, NH ₂ , NO ₂ or CN, in particular H, Cl, OH, CH ₃ or NH ₂
Ar phenyl
mean;
in Id R ¹ H, OH, OA, SA or Hal, in particular H, OH, OCH ₃ or CH ₃
R ² H, Hal, OH, A, NH ₂ , NO ₂ or CN, in particular H, Cl, OH, CH ₃ or NH ₂
Ar phenyl
R ^{3 is} H, A, NH ₂ or SA, especially H or CH ₃ ;
in Ie R ¹ H, OH, OA, SA or Hal, in particular H, OH, OCH ₃ or CH ₃
R ² H, Hal, OH, A, NH ₂ , NO ₂ or CN, in particular H, Cl, OH, CH ₃ or NH ₂
Ar phenyl
R ^{3 is} H, A, NH ₂ or SA, in particular H or CH ₃
R ^{4 is} H, Hal, NH ₂ , or NO _2, in particular H or NH ₂ .

Also preferred are those compounds of the formula I and their salts and solvates in which R simultaneously denotes H, Ar phenyl and at least one of the radicals R ¹ , R ² , R ³ , R ^{4 has} one of the following meanings:
OH, NO ₂ , NH ₂ , NHA, NA ₂ , NH-CO-A, NH-CO-Ph, SA, SO-A, SO ₂ -A, SO ₂ -Ph, CN, OCF ₃ , CO-A, CO ₂ H, CO ₂ A, CO-NH ₂ , CO-NHA, CO-NA ₂ , SO ₂ NH ₂ , SO ₂ NHA, SO ₂ NA ₂ or unsubstituted or singly or multiply by A, OA, Hal, CF ₃ substituted phenyl. Of these compounds, those are particularly preferred whose radical R ^{1 is} Cl, in particular in position 6, and those compounds whose radical R ^{3 is} methyl, in particular in position 4 '.

Also preferred are those compounds of the formula I and their salts and solvates in which the radicals R ⁵ , R ⁶ , R ⁷ , R ⁸ have the meaning H at the same time. Of these compounds, those are particularly preferred whose radical R ^{1 is} Cl, in particular in position 6, and those compounds whose radical R ^{3 is} methyl, in particular in position 4 ', and those compounds whose radical R ^{4 is} NH ₂ , in particular in position 2 ', means.

Compounds of the formula I, the radical R ^{3 of which is} methyl, in particular in the 4 'position, have a particularly pronounced selectivity of the binding to the NHE-3 receptor.

Compounds of the formula I, the radical R ^{4 of which is} NH ₂ , in particular in the position 2 ', show particularly good solubility in aqueous solutions.

Compounds of the formula I in which R ^{1 is} H, R ² Cl in the 6-position and R ^{3 is} methyl in the 4'-position are preferred. Compounds of the formula I are very particularly preferred, the radical R ^{4 of which} additionally denotes NH ₂ in the 2'-position.

The compounds of the formulas If to Ik are particularly preferred:

wherein R ¹ , R ² , R ³ , R ⁴ and Y have the meaning given above and R ^{1 is} preferably H, OH, OA, SA, or F, in particular H, OH, OCH ₃ or CH ₃ . R ¹ in the formulas If to Ik very particularly preferably denotes H.

R ² preferably denotes H, Cl, A, NH ₂ , NO ₂ , SCH ₃ , SOCH ₃ , SO ₂ CH ₃ , OCH ₃ , OH, CN, CF ₃ , OCF ₃ or F, in particular H, Cl, F, Br , OH, CH ₃ , NO ₂ or NH ₂ , R ² in the formulas If to Ik very particularly preferably denotes Cl.

R ³ preferably denotes H, Cl, A, NH ₂ , NO ₂ , SCH ₃ , CN, C ₂ H ₅ , OCF ₃ or C ₆ H ₅ , in particular H, A or CH ₃ . R ³ in the formulas If to Ik very particularly preferably denotes CH ₃ .

R ⁴ preferably denotes H, F, NH ₂ or NO ₂ , in particular H or NH ₂ . R ⁴ in the formulas If to Ik very particularly preferably denotes NH ₂ .

Y has the meaning given above in the formulas If to Ik. Y preferably has one of the following meanings:

Y particularly preferably has one of the following meanings:

The following compounds I1 to I10 and their salts and solvates are also particularly preferred:

N- (6-chloro-4-phenyl-quinazolin-2-yl) -N'-methyl-guanidine I1

N- (6-chloro-4-p-tolyl-quinazolin-2-yl) -N'-methyl-guanidine I2

N- [6-chloro-4- (2-nitro-phenyl) -quinazolin-2-yl] -N'-methyl-guanidine I3

N- [4- (2-Amino-phenyl) -6-chloro-quinazolin-2-yl] -N'-methyl-guanidine 14

N- [6-Chloro-4- (4-methyl-2-nitro-phenyl) -quinazolin-2-yl] -N'-methyl-guanidine I5

N- [4- (2-Amino-4-methylphenyl) -6-chloro-quinazolin-2-yl] -N'-methyl-guanidine I6

N- [6-Chloro-4- (2-nitro-phenyl) -quinazolin-2-yl] guanidine I7

N- [4- (2-Amino-phenyl) -6-chloro-quinazolin-2-yl] guanidine I8

N- [6-Chloro-4- (4-methyl-2-nitro-phenyl) -quinazolin-2-yl] guanidine I9

N- [4- (2-Amino-4-methyl-phenyl) -6-chloro-quinazolin-2-yl] - guanidine I10

The hydrochlorides and p-toluenesulfonates of the compounds of the formulas I1 to I10 are very particularly preferred.

The compounds of formula I and also the starting materials for their manufacture position are otherwise produced by methods known per se, as described in literature (e.g. in standard works such as Houben-Weyl, Methods of Organic Chemistry, Georg-Thieme-Verlag, Stuttgart) be are written, namely under reaction conditions for the ge  mentioned implementations are known and suitable. You can also do that use of known variants not mentioned here do.

If desired, the starting materials can also be formed in situ, so that they are not isolated from the reaction mixture, but immediately further reacted to the compounds of formula I.

The 2-guanidino-4-aryl-quinazolines of the formula I are preferably prepared by using o-aminophenyl ketones or o-aminonaphthyl ketones of the formula II

wherein R ¹ , R ² and Ar have the meanings given in claim 1, with 1-cyanguanidine or a correspondingly N-alkylated or N-arylated 1-cyanguanidine of the formula NC-Y, in which Y has the meaning given above.

The reaction can be carried out in an inert solvent.

Suitable inert solvents are, for. B. hydrocarbons such as hexane, Petroleum ether, benzene, toluene or xylene; chlorinated hydrocarbons such as Trichlorethylene, 1,2-dichloroethane, carbon tetrachloride, chloroform or dichloromethane; Alcohols such as methanol, ethanol, isopropanol, n-propanol, n-butanol or tert-butanol; Ethers such as diethyl ether, diisopropyl ether, Te trahydrofuran (THF) or dioxane; Glycol ethers such as ethylene glycol mono methyl or monoethyl ether (methyl glycol or ethyl glycol), ethylene glycol dimethyl ether (diglyme); Ketones such as acetone or butanone; amides such as acetamide, dimethylacetamide, N-methylpyrrolidone (NMP) or dime thylformamide (DMF); Nitriles such as acetonitrile; Sulfoxides such as dimethyl sulfoxide (DMSO); Carbon disulphide; Carboxylic acids such as formic acid or Es  sigsäure; Nitro compounds such as nitromethane or nitrobenzene; Esters like Ethyl acetate or mixtures of the solvents mentioned.

DMF, water or an alcohol is preferably used. The reaction is very particularly preferably without a solvent, i. H. in the melt, at temperatures between 100 and 200 ° C leads.

The presence of an acidic catalyst such as AlCl ₃ , TiCl ₄ , p-toluenesulfonic acid, BF ₃ , acetic acid, sulfuric acid, oxalic acid, POCl ₃ or phosphorus pentoxide is advantageous.

A preferred variant is that one of the reactants already as salt, e.g. B. is used as the hydrochloride.

Another valuable method for preparing the compounds of formula I is that instead of a compound of formula NC-Y, a compound of formula III

HN = CX-Y III

wherein
X -S-alkyl, -S-aryl, -O-alkyl or -OAryl
and alkyl preferably has the meaning of A given above and aryl has the meaning of Ar given above,
reacted with a compound of formula II.

Finally, the compounds of formula I by reacting 2-chloro-4-arylquinazolines of formula IV

wherein Ar, R ¹ and R ^{2 have} the meanings given above,
be prepared with a compound of formula HY, wherein Y has the meaning given above. HY is particularly preferably guanidine.

A base of formula I can with an acid in the associated acid addition salt can be transferred, for example by reaction equi valent amounts of the base and the acid in an inert solvent such as ethanol and subsequent evaporation. For this implementation men in particular acids, the physiologically acceptable Sal ze deliver. So inorganic acids can be used, e.g. B. Sulfuric acid, nitric acid, hydrohalic acids such as chlorwas hydrochloric acid or hydrobromic acid, phosphoric acids such as ortho phosphoric acid, sulfamic acid, also organic acids, in particular aliphatic, alicyclic, araliphatic, aromatic or heterocyclic mono- or polybasic carboxylic, sulfonic or sulfuric acids, e.g. B. Amei sensic acid, acetic acid, propionic acid, pivalic acid, diethyl acetic acid, Malonic acid, succinic acid, pimelic acid, fumaric acid, maleic acid, Lactic acid, tartaric acid, malic acid, citric acid, gluconic acid, Ascor bic acid, nicotinic acid, isonicotinic acid, methane or ethanesulfonic acid, Ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, p- Toluenesulfonic acid, naphthalene mono- and disulfonic acids, lauryl sulfur acid. Salts with physiologically unacceptable acids, e.g. B. picrates, can for the isolation and / or purification of the compounds of For mel I can be used.

The invention further relates to the use of the compounds of the formula I as NHE-3 inhibitors and / or their physiologically harmless Lichen salts for the production of pharmaceutical preparations, in particular the non-chemical way. Here you can together with min at least a solid, liquid and / or semi-liquid carrier or auxiliary fabric and possibly in combination with one or more width ren active ingredients are brought into a suitable dosage form.

The invention furthermore relates to pharmaceutical preparations, containing at least one NHE-3 inhibitor of the formula I and / or one its physiologically acceptable salts and solvates.  

These preparations can be used as medicinal products in human or veteri used in medicine. Organic or inorganic substances in question that are suitable for enteral (e.g. oral), parenteral or topical application and with the new compound not react, e.g. water, vegetable oils, benzyl alcohols, alkylene glycols, polyethylene glycols, glycerol triacetate, gelatin, Carbohydrates such as lactose or starch, magnesium stearate, talc, petroleum jelly. Tablets, pills, coated tablets, Capsules, powder, granules, syrups, juices or drops, for rectal application application suppositories, for parenteral application solutions, preferred wise oily or aqueous solutions, also suspensions, emulsions or implants, for topical application of ointments, creams or powder, or transdermally in patches.

The new compounds can also be lyophilized and the Lyo obtained philisate z. B. can be used for the preparation of injectables. The specified preparations can be sterilized and / or auxiliary substances such as lubricants, preservatives, stabilizers and / or wetting agents, Emulsifiers, salts to influence the osmotic pressure, buffers substances, colors, flavors and / or several other active ingredients included, e.g. B. one or more vitamins.

As a pharmaceutical preparation for administration in the form of aero brines or sprays are suitable e.g. B. solutions, suspensions or emul Sions of the active ingredient of formula I in a pharmaceutically acceptable Chen solvent.

The compounds of formula I and their physiologically acceptable Salts and solvates can be used to treat and / or prevent the above described diseases or disease states can be used.

The substances according to the invention are generally preferred as in dosages between about 0.1 and 100 mg, in particular between between 1 and 10 mg per dosage unit. The daily dose tion is preferably between about 0.001 and 10 mg / kg body weight. However, the specific dose for each patient depends on the different most factors, for example the effectiveness of the used  special connection, age, body weight, general health condition, gender, diet, time of administration and path, the rate of excretion, drug combination and Severity of the respective disease to which the therapy applies. The oral Application is preferred.

Examples example 1

A mixture of 1.00 g of 2-amino-5-chloro-2'-nitrobenzophenone, 0.60 g of 1- Cyanguanidine and 2.00 g of p-toluenesulfonic acid monohydrate were added for 2 h melted at 150 ° C. The cooled melt was mixed with methanol sets and for 30 min. stirred at 65 ° C. The return obtained after filtration was discarded and water was added to the filtrate. Subsequently the solution was made alkaline and extracted with ethyl acetate. The ex tract was concentrated and crystallized from acetonitrile, thereby freeing Base N- [6-chloro-4- (2-nitro-phenyl) -quinazolin-2-yl] guanidine was obtained de.

To form the acid addition salt, the base was dissolved in methanol, the mixture is acidified with HOI-containing isopropanol and the solution medium then removed. Crystals of N- [6-chloro-4- (2-nitro-phenyl) - quinazolin-2-yl] guanidinium chloride could be obtained from acetonitrile the.

Example 2

1.20 g of N- (5-methoxy-4-phenyl-quinazolin-2-yl) guanidinium chloride were stirred with 8.00 g of pyridinium chloride at 170 ° C. for 6 h. The cooled melt was then treated with 20 ml of a Na ₂ S ₂ O ₄ solution. The precipitate obtained was isolated, dissolved in methanol and the solution acidified with HCl-containing isopropanol. After removal of the solution by means of, the residue was crystallized from acetonitrile, giving N- (5-hydroxy-4-phenyl-quinazolin-2-yl) guanidinium chloride (mp. 310 ° C.).

Example 3

A mixture of 3.01 g of 2-amino-5-chlorobenzophenone, 2.55 g of N-cyano N'-methylguanidine and 7.42 g of p-toluenesulfonic acid monohydrate was used for Stirred for 2 h at 150 to 160 ° C in the melt. The cooled melt was mixed with methanol and for 30 min. stirred at 65 ° C. The after Filtration residue was discarded, the filtrate with water and Ethyl acetate was added and again for 30 min. stirred at 65 ° C. Subsequently was allowed to crystallize with stirring in an ice bath, whereby N- (6-chloro-4- phenyl-quinazolin-2-yl) -N'-methyl-guanidinium-p-toluenesulfonate obtained (mp 268-269 ° C).

Example 4

300 mg of N- [6-chloro-4- (2-nitro-phenyl) -quinazolin-2-yl] guanidinium-p- Toluene sulfonate was dissolved in 50 ml of methanol and in the presence of 300 mg Raney nickel at RT within 21 h under normal pressure hydrogenated. After filtration and removal of the solvent, N- [6- Chloro-4- (2-aminophenyl) quinazolin-2-yl] guanidinium p-toluenesulfonate the filtrate. (Mp 250 ° C).

Example 5

A mixture of 0.350 g of N- (6-methylsulfanyl-4-phenyl-quinazolin-2-yl) - guanidinium chloride and 0.140 g sodium perborate trihydrate in 5 ml vinegar acid was for 30 min. stirred at 80 ° C. Then the Lö concentrated and mixed with water. The aqueous solution was on Adjusted pH 12 and extracted with ethyl acetate. By narrowing the ex tracts gave N- (6-methanesulfinyl-4-phenyl-quinazolin-2-yl) guanidine in crystalline form (mp. 175-180 ° C).

Example 6

A mixture of 1,200 g of N- (6-methylsulfanyl-4-phenyl-quinazolin-2-yl) - guanidinium chloride and 0.154 g sodium perborate trihydrate in 5 ml vinegar acid was stirred at 80 ° C for 1 h. Then the reaction the mixture was concentrated and water was added. The solution obtained was adjusted to pH 12 and extracted with ethyl acetate. By narrowing the Extract was obtained N- (6-methanesulfonyl-4-phenyl-quinazolin-2-yl) - guanidine in crystalline form (mp. 180-185 ° C).  

To form the acid addition salt, 0.80 g of N- (6-methanesulfonyl- 4-phenyl-quinazolin-2-yl) guanidine with an aqueous 1 N HCl solution acts and the crystals obtained recrystallized from ethanol.

Example 7

2.70 g of the hydrochloride of 2-amino-5-chlorobenzophenone and 1.70 g of N- Cyan-N ', N "-dimethyl-guanidine were mixed and he at 150 ° C for 3 h hitzt. The reaction product was taken up in methanol and filtered. The filtrate was restricted. The residue was made up of a mixture of Isopropanol and diethyl ether recrystallized, whereby N- (6-chloro-4-phenyl quinazolin-2-yl) -N ', N "-dimethyl-guanidinium chloride was obtained (mp. 264-267 ° C).

Example 8

A mixture of 500 mg of 2-amino-5-chloro-2'-nitrobenzophenone, 406 mg N-cyan-N-ethylguanidine and 1.03 g of p-toluenesulfonic acid monohydrate were de stirred for 2 h at 150 to 160 ° C in the melt and as in Example 3 worked up, whereby N- [6-chloro-4- (2-nitro-phenyl) -quinazolin-2-yl] -N'- ethyl guanidinium p-toluenesulfonate was obtained (mp. 298-300 ° C).

Example 9

A mixture of 500 mg of 2-amino-5-chloro-2'-nitrobenzophenone, 580 mg N-cyan-N-phenylguanidine and 1.03 g of p-toluenesulfonic acid monohydrate was stirred for 2 h at 150 to 160 ° C in the melt and as in Bei game 3 worked up, whereby N- [6-chloro-4- (2-nitro-phenyl) -quinazoline-2- yl] -N'-phenyl-guanidinium p-toluenesulfonate was obtained (mp. 261-263 ° C).

Analogously to the above-mentioned methods, the following were obtained as preferred NHE-3 inhibitors using the corresponding precursors: acid addition salts:
In the following, pTsOH means p-toluenesulfonic acid.

Examples 10-94

Examples 95-147

Examples 148-198

Examples 199-279

Examples 280-364

Examples 365-449

Examples 450-534

Examples 535-619

Examples 620-704

Examples 705-789

Examples 790-874

Pharmacological tests

The following describes the methodology used to characterize the Compounds of formula I was used as NHE-3 inhibitors.

The compounds of formula I were characterized in terms of their selectivity against the isoforms NHE-1 to NHE-3. The three isoforms were stably expressed in mouse fibroblast cell lines. The inhibitory effect of the compounds was assessed by determining the EIPA-sensitive ²² Na ⁺ uptake into the cells after intracellular acidosis.

To characterize the Na ⁺ / H ⁺ exchange inhibitors with regard to their isoform selectivity, we examined the compounds for their inhibition of the NHE isoforms NHE-1, -2 and -3, which were stably expressed in a mouse fibroblast cell line (see process part), since by that the EIPA-sensitive ²² Na ⁺ uptake into the cells was determined after in tracellular acidosis.

material and methods LAP1 cell lines that express the different NHE isoforms

The LAP1 cell lines expressing the isoforms NHE-1, -2 and -3 (a mouse fibroblast cell line) were obtained from Prof. J. Pouysségur (Nice, Frank Reich). The transfections were carried out according to the method of Fran chi et al. (1986). The cells were cultured in Dulbecco's modified Eagle medium (DMEM) with 10% inactivated fetal calf serum (FCS). For the selection of the NHE-expressing cells, the so-called "acid killing method" by Sardet et al. (1989) used. The cells were first removed in a NH ₄ Cl-containing bicarbonate and sodium-free buffer by washing with a bicarbonate, NH ₄ Cl and sodium-free buffer and incubated with a bicarbonate-free NaCl-containing buffer. Only those cells that functionally express NHE survived in the intracellular acidification to which they were exposed.

Characterization of NHE inhibitors in relation to their isoform selects tivity

With the above mouse fibroblast cell lines expressing the isoforms NHE-1, NHE-2 and NHE-3, compounds according to the method described by Counillon et al. (1993) and Scholz et al. (1995) described procedure for selectivity compared to the isoforms. The cells were acidified intracellularly using the NH ₄ Cl prepulse method and then by incubation in a bicarbonate-free ²² Na ⁺ -containing buffer. Due to the intracellular acidification, NHE was activated and sodium was absorbed into the cells. The effect of the test compound was expressed as an inhibition of EIPA (ethyl isopropylamiloride) sensitive ²² Na ⁺ uptake.

The cells expressing NHE-1, NHE-2 and NHE-3 were inoculated at a density of 5-7.5 x 10 ⁴ cells / well in microtiter plates with 24 wells and grown to confluence for 24 to 48 hours. The medium was aspirated and the cells were incubated for 60 minutes at 37 ° C. in the NH ₄ Cl buffer (50 mM NH ₄ Cl, 70 mM choline chloride, 15 mM MOPS, pH 7.0). The buffer was then removed and the cells were rapidly overlaid twice with the choline chloride washing buffer (120 mM choline chloride, 15 mM PIPES / Tris, 0.1 mM ouabain, 1 mM MgCl ₂ , 2 mM CaCl ₂ , pH 7.4); the cells were incubated in this buffer for 6 minutes. After the incubation period had elapsed, the incubation buffer was aspirated. To remove extracellular radioactivity, the cells were quickly washed four times with ice-cold phosphate-buffered saline (PBS). The cells were then solubilized by adding 0.3 ml of 0.1 N NaOH per well. The cell fragment-containing solutions were transferred to scintillation tubes. Each well was washed twice more with 0.3 ml of 0.1 N NaOH and the washing solutions were also added to the corresponding scintillation tubes. The tubes containing the cell lysate were mixed with scintillation cocktail and the radioactivity absorbed into the cells was determined by determining the β radiation.

Inhibition of ²² Na ⁺ uptake in rabbit erythrocytes

Na ⁺ / H ⁺ exchange activity was also determined by observing the uptake of ²² Na ⁺ ions in acidified rabbit erythrocytes. Rabbit erythrocytes have found widespread use in studies of Na ⁺ / H ⁺ exchange activity (Escobales & Fugueroa, 1991; Morgan & Canessa, 1990). The EIPA-sensitive portion of the ²² Na ⁺ uptake in acidified erythrocytes was regarded as Na ⁺ / H ⁺ -dependent Na ⁺ uptake.

cell preparation

Red blood cell preparation and internal acidification The red blood cells were strongly based on the procedures by Morgan and Canessa (1990).

The blood was obtained from rabbits (e.g. New Zealand White). It was collected in 50 ml Falcon centrifuge tubes containing 5 ml of sodium heparin solution (250 U / ml). The blood and heparin solution were mixed well. The red blood cells were obtained by centrifugation at 2000 × g at 4 ° C; Plasma and white blood cell cuff were removed. The remaining solution was filtered through 200 µm gauze. The filtrate was resuspended with wash buffer to the original volume (140 mM KCl, 0.15 mM MgCl ₂ , 10 mM TRIS / MOPS, pH 7.4). The red blood cells were again obtained by centrifugation (2000 × g, 4 ° C.). The washing process was repeated twice.

Intracellular acidification

For intracellular acidification, 5 ml of the deposited collected red blood cells were again suspended with 45 ml acidification buffer (170 mM KCl, 0.15 mM MgCl ₂ , 0.1 mM ouabain, 10 mM glucose, 10 mM sucrose, 20 mM Tris / Mes, pH 6.2). The red blood cell suspension was incubated for 10 minutes at 37 ° C (with occasional mixing). To fix the internal pH, up to 200 µM and 1 mM DIDS or DIAMOX (acetazolamide) are added. Incubation was continued for 30 minutes at 37 ° C.

The red blood cells were then collected by centrifugation (4 minutes at 2000 × g, 4 ° C); they were again suspended with ice-cold unbuffered washing solution (170 mM KCl, 40 mM sucrose, 0.15 mM MgCl ₂ ) and thus washed four times.

Incubation and measurement of the ²² Na ⁺ uptake

The incubation was carried out in Macrowell tube strips in a format of 8 × 12. The incubation was started by adding 200 .mu.l incubation buffer (160 mM KCl, ²² NaCl (37 MBq / well), 10 mM NaCl, 0.15 mM MgCl ₂ , 0.1 mM ouabain, 10 mM glucose, 40 mM Saccha rose, 10 mM Tris / MOPS, pH 8.0, 0.5 mM Diamox, 1% DMSO) with 20 μl of the (preheated) acidified solution of the red blood cells. The test substances were first dissolved with 100% DMSO and the solution was then diluted to the appropriate concentrations with incubation buffer. The mixture was incubated at 37 ° C. for 5 minutes. (In preliminary experiments it was shown that under these incubation conditions the ²² Na uptake rate was linear during the 5 minute incubation period). The incubation was stopped by adding 800 μl of ice-cold stop solution (112 mM MgCl ₂ , 0.1 mM ouabain). The tubes were kept on ice for a short time. The tubes were then covered with parafilm and the red blood cells were obtained by centrifugation at 2000 × g and 4 ° C. for 7 minutes. The supernatant was aspirated with a self-made suction device, with which one could aspirate 4 adjacent tubes at the same time; Spacer rings on the further ends of the tips prevented the tips from penetrating too deeply into the tubes and the pellet of the red blood cells from being aspirated. All ²² Na-containing supernatants and washing solutions were kept and disposed of as radioactive waste.

The red blood cells were three times with 900 ul ice-cold stop solution washed, and that by the Sus oscillation / centrifugation step repeated. After the last wa  The pellet of the red blood cells was washed with 200 µl water puts. The tubes were then sonicated 2 × 30 minutes treated. The Macrowell tube strips were then separated and each tube was turned upside down into its own scintile given lation tubes; the shake emptied by gentle shaking Molysed solution of the red blood cells in the scintillation vial. Each vial was washed with 3 ml of Aquasafe 300 scintillation fluid PS offset, and the jars were sealed and good mixed.

The radioactivity absorbed into the red blood cells was determined in an egg nem scintillation counter determined by tracking the β decay.

The determination was carried out in triplicate for each substance concentration. The mean of the count was presently subtracted from each value of 10 µM EIPA to include the non-Na ⁺ / H ⁺ -dependent ²² Na ⁺ uptake in the erythrocytes. The mean of the remaining counts in the absence of a substance was used as a 100% control; the mean values in the presence of the test compounds were expressed as a percentage of this control value. The percentages of the recording data were recorded semi-logarithmically; IC50 values were obtained by fitting the values to a nonlinear curve using the equation f (x) = 100 / (1 + (IC50 / x) ** n).

literature

Counillon et al. (1993) Mol. Pharmacol. 44: 1041-1045
Escobales and Figueroa (1991) J. Membrane Biol. 120, 41-49
Franchi et al. (1986) Proc. Natl. Acad. Sci. USA 83: 9388-9392
Morgan and Canessa (1990) J. Membrane Biol. 118, 193-214
Sardet et al. (1989) Cell 56: 271-280
Scholz et al. (1995) Cardiovasc. Res. 29: 260-268.

The following examples relate to pharmaceutical preparations:

Example A injection Vials

A solution of 100 g of an NHE-3 inhibitor of formula I and 5 g of Dina trium hydrogen phosphate is in 3 l of double distilled water with 2 n Hydrochloric acid adjusted to pH 6.5, sterile filtered, filled into injection glasses, lyophilized under sterile conditions and sealed sterile. Every In jection glass contains 5 mg of active ingredient.

Example B suppositories

A mixture of 20 g of an NHE-3 inhibitor of the formula I is melted with 100 g soy lecithin and 1400 g cocoa butter, pour into molds and leave cool. Each suppository contains 20 mg of active ingredient.

Example C solution

A solution is prepared from 1 g of an NHE-3 inhibitor of the formula I, 9.38 g of NaH ₂ PO ₄ .2 H ₂ O, 28.48 g of Na ₂ HPO ₄ .12 H ₂ O and 0.1 g of benzalko nium chloride in 940 ml of double distilled water. It is adjusted to pH 6.8, made up to 1 l and sterilized by irradiation. This solution can be used in the form of eye drops.

Example D ointment

500 mg of an NHE-3 inhibitor of the formula I are mixed with 99.5 g of Vaseli ne under aseptic conditions.

Example E tablets

A mixture of 1 kg of an NHE-3 inhibitor of the formula I, 4 kg of lactose, 1.2 kg of potato starch, 0.2 kg of talc and 0.1 kg of magnesium stearate is in Compressed into tablets in the usual way, such that each tablet 10 mg Contains active ingredient.  

Example F dragees

Analogously to Example E, tablets are pressed, which are then made in the usual manner Wise with a coating of sucrose, potato starch, talc, tragacanth and dye are coated.

Example 6 capsules

2 kg of an NHE-3 inhibitor of the formula I are hard in the usual way filled gelatin capsules so that each capsule contains 20 mg of the active ingredient.

Example H ampoules

A solution of 1 kg of NHE-3 inhibitor of the formula I in 60 l of double distillation filtered water is sterile filtered, filled into ampoules, under sterile loading conditions lyophilized and sealed sterile. Each ampoule contains 10 mg of active ingredient.

Claims (13)

1. Compounds of formula I.
Ar unsubstituted or simply substituted by R ³ and / or R ⁴ phenyl or naphthyl,
R ¹ , R ² , R ³ , R ⁴ each independently of one another H, A, OA, Hal, CF ₃ , OH, NO ₂ , NH ₂ , NHA, NA ₂ , NH-CO-A, NH-CO-Ph, SA, SO-A, SO ₂ -A, SO ₂ -Ph, CN, OCF ₃ , CO-A, CO ₂ H, CO ₂ A, CO-NH ₂ , CO-NHA, CO-NA ₂ , SO ₂ NH ₂ , SO ₂ NHA, SO ₂ NA ₂ or unsubstituted or mono- or polysubstituted by A, OA, Hal, CF ₃
A alkyl having 1, 2, 3, 4, 5 or 6 carbon atoms,
Hal F, Cl, Br or I
R ⁵ , R ⁶ , R ⁷ , R ⁸ each independently represent H, A, unsubstituted or mono- or polysubstituted by A, OA, Hal, CF ₃ , where R ⁵ and R ⁷ , R ⁵ and R ⁶ , R ⁷ and R ^{8 can form} 5-7-membered rings,
and their salts and solvates, with the proviso that compounds in which R ⁵ , R ⁶ , R ⁷ and R ^{8 have} the meaning H and none of the radicals R ¹ , R ² , R ³ , R ⁴ OH, NO ₂ , NH ₂ , NHA, NA ₂ , NH-CO-A, NH-CO-Ph, SA, SO-A, SO ₂ -A, SO ₂ -Ph, CN, OCF ₃ , CO-A, CO ₂ H, CO ₂ A, CO-NH ₂ , CO-NHA, CO-NA ₂ , SO ₂ NH ₂ , SO ₂ NHA, SO ₂ NA ₂ or unsubstituted or mono- or polysubstituted by A, OA, Hal or CF ₃ , with exception of. 2. Compounds of formula I according to claim 1 and their salts and Solvates as NHE 3 inhibitors. 3. Compounds of formula I according to claim 1 and their physiological harmless salts or solvates for use in loading fighting diseases. 4. Use of compounds of formula I according to claim 1 and / or their physiologically acceptable salts or solvates for Manufacture of a drug. 5. Use of compounds of formula I according to claim 1 and ih re physiologically acceptable salts and / or solvates provision of a medicament for the treatment of hypertension, from Thrombosis, ischemic conditions of the heart, peripheral and central nervous system and stroke, ischemic Conditions of peripheral organs and limbs and for treatment of shock conditions. 6. Use of compounds of formula I according to claim 1 and ih re physiologically acceptable salts and / or solvates Provision of a drug for use in surgical operations  organ transplants and for preservation and location grafts for surgical measures. 7. Use of compounds of formula I according to claim 1 and ih re physiologically acceptable salts and / or solvates provision of a medicinal product for the treatment of diseases in which de cell proliferation is a primary or secondary cause provides, for the treatment or prophylaxis of disorders of the fatty substance alternating or disturbed breath drive. 8. Use of compounds of formula I according to claim 1 and ih re physiologically acceptable salts and / or solvates provision of a medicinal product for the treatment of ischemic kidney, ischemic bowel diseases or for the prophylaxis of acute or chronic kidney disease. 9. Use of compounds of formula I according to claim 1 and ih re physiologically acceptable salts and / or solvates provision of a drug for the treatment of bacterial and pa racy diseases. 10. Pharmaceutical preparation, characterized by a content at least one NHE-3 inhibitor according to claim 1 and / or one their physiologically acceptable salts and / or solvates. 11. Compounds selected from the group of compounds I1 to I10
N- (6-chloro-4-phenyl-quinazolin-2-yl) -N'-methyl-guanidine I1
N- (6-chloro-4-p-tolyl-quinazolin-2-yl) -N'-methyl-guanidine I2
N- [6-chloro-4- (2-nitro-phenyl) -quinazolin-2-yl] -N'-methyl guanidine I3
N- [4- (2-Amino-phenyl) -6-chloro-quinazolin-2-yl] -N'-methyl guanidine 14
N- [6-Chloro-4- (4-methyl-2-nitro-phenyl) -quinazolin-2-yl] -N'-methyl-guanidine I5
N- [4- (2-Amino-4-methylphenyl) -6-chloro-quinazolin-2-yl] -N'-methyl-guanidine I6
N- [6-Chloro-4- (2-nitro-phenyl) -quinazolin-2-yl] guanidine I7
N- [4- (2-Amino-phenyl) -6-chloro-quinazolin-2-yl] guanidine I8
N- [6-Chloro-4- (4-methyl-2-nitro-phenyl) -quinazolin-2-yl] guanidine I9
N- [4- (2-Amino-4-methylphenyl) -6-chloroquinazolin-2-yl] guanidine I10
and their salts and solvates. 12. Compounds according to claim 1 as active pharmaceutical ingredients. 13. A process for the preparation of the 2-guanidino-4-aryl-quinazolines of the formula I and their salts and solvates, characterized in that either

• a) Compounds of formula II
  wherein R ¹ , R ² and Ar have the meanings given above, with 1-cyanguanidine or a correspondingly N-alkylated or N-arylated cyanguanidine of the formula NC-Y, wherein Y has the meaning given in claim 1

or

• a) instead of a compound of formula NC-Y a compound of formula III
  HN = CX-Y III
  wherein X is -S-alkyl, -S-aryl, -O-alkyl or -OAryl, reacted with a compound of formula II

or

• a) 2-chloro-4-arylquinazolines of the formula IV
  wherein Ar, R ¹ and R ^{2 have} the meanings given in claim 1 with
  a compound of formula HY, wherein Y has the meaning given in claim 1
  and optionally following steps (a), (b) or (c) converts a basic or acidic compound of the formula I into one of its salts or solvates by treatment with an acid or base.